Quality Control School

Yaping Gu

**** *. ******* ***., ***. ***

Chicago, IL 60615

Cell Phone 440-***-****

E-mail: *****@*****.***

OBJECTIVE

To obtain a position allowing me to utilize my knowledge and skills in the field of molecular and cell biology, immunology, biochemistry, and animal study

SUMMARY

• Educational Qualifications:

- M.S. in Pathophysiology (University of Suzhou, School of Medicine)

- M.D. in Medicine (University of Nantong, School of Medicine)

- Scholar in Molecular Cardiovascular Medicine (University of Tokyo, School of Medicine)

- Fellow in Molecular & Cellular Biology ( Case Western Reserve University, School of Medicine)

• Research Experience:

- Extensive experience in the field of molecular & cell Biology, biochemistry, and genetics, pathophysiology, and animal model of diseases.

- Accomplished research projects related to neuron degeneration disease, growth hormone signal transduction pathway, ischemic-reperfusion injury and apoptosis.

• Industry Experience:

- Ion channel gene cloning, mammalian stable cell line development and validation.

- Application of Fluorometric Imaging Plate Reader ( FLIPR)and FLEX station in cell-based ion channel screens & concentration response assays for cardiac safety and drug discovery.

• Academic and Scientific Distinctions:

- Received national and international awards and fellowships.

- Obtained prizes for excellent teaching and research.

- Published 32 papers in peer-reviewed Journals.

- Reviewer of Jiaotong Yixue, and Acta Academiae Med. Nantong.

• Communication/Presentation skills:

- Ability in communication with students, colleagues, supervisors, by which built good relationship and work environment.

- Annually presenting research achievements in national and international conferences organized by American Society for Cell Biology and American Society for Neurosciences. Japenese Society of Circulation, Chinese Society of Pathophysiology

• Computer Software:

- Microsoft Word, Excel, and Power Point, Adobe Photoshop, Adobe Illustrator

EMPLOYMENT HISTORY

2006-present: Research Scientist. ChanTest Corp

• Ion channel gene cloning: Applied molecular biology techniques to clone various ion channels

including voltage-gated or ligand-gated potassium, sodium, and calcium channels to expression

vectors.

• Cell line development and validation: Transiently or stably expressed ion channel gene in HEK and

CHO cells. Did the characterization of cellular processed protein and functional validation.

• Developed cell based ion channel screens & concentration response assays by for drug discovery

and cardiac safety channel panel screening.

2004-2006: Senior Research Associate. Institute of Pathology, Case Western Reserve Univ

• Developed and studied cell models of prion protein (PrP) insertion and deletion.

• Prepared small grant proposal ‘Pathogenic mechanisms in inherited prion disorders’.

• Studied the rout of PrP scrapie transmission.

1999-2003: Research Associate. Institute of Pathology, Case Western Reserve Univ

• Studied nucleation role of PrP on the cell model treated with toxic peptide.

• Proved that high level of CTM PrP is induced by PrP aggregation.

• Demonstrated the existence of nuclear localization signal in prion N-terminus that triggers nuclear

transportation when prion becomes C-terminal truncated form.

• Studied the effect of protein-folding agents on a cell model of CJD H187R and indicate the

Important role of ER quality control in phenotype-rescue .

1997-1999: Research Scholar, Univ of Tokyo School of Medicine

• Identified growth hormone signaling pathway leading to ERK activation in cardiac myocytes and the suppression of growth hormone on hydrogen peroxide-induced apoptosis in cardiomyocytes.

• Reported the cardiac protective effect of Insulin from oxidative stress-induced apoptosis through

• activation of PI3 Kinase/Akt.

• Collaboratively studied on the role of calcium in Angiotensin II-induced hypertrophy.

1994-1996: Assistant Professor, Dept of Pathophysiology, Univ of Nantong School of medicine

• As principal investigator for the project ‘Mechanisms of myocardioum ischemic-reperfusion injury’ supported by Nantong Science and Technology Committee, China.

• Examined the effect of ROS on sulfhydryl groups and Ca2+-pump activity of cardiac myosin.

• Studied the role of heat shock protein in preconditioning myocardium.

1988-1993: Instructor, Dept of Pathophysiology, Univ of Nantong School of medicine

Focused on ‘the mechanisms of occupational diseases of the oil vessel-working members’ sponsored by Chinese Ministry of Communications.

• Revealed that the lipid peroxidation caused by the inhalation of crude petroleum vapor was

involved in pathogenesis of rat lung dysfunction

• Developed the method of Chromotest and examined the toxicity of crude oil in mutagenesis.

• Found that the inhalation of petroleum vapor changes the level of neuronal transmitters in rat

brain by employing HPLC instrument to this study.

1984-1987: Graduate Student, Dept of Pathophysiology, Univ. of Shuzou School of Medcine

Studied the pathophysiological mechanisms of shock:

• Did the quantification study on the generation of reactive oxygen species in plasma and liver

of rat and dog after subjected to the hemorrhagic shock.

• Examined the protective role of superoxide dismutase from free radical injury by monitoring

the level of hepatic lysosome enzyme released in hemorrhage-shocked rat.

LABORATORY TECHNIQUES PROFICIENCY

• Molecular biology:

- Gene cloning related techniques: RT-PCR for Synthesis of cDNA from different tissue mRNA. Design oligo primers for PCR. Bacteria transformation and positive clone screening. Manual and automation isolation of DNA plasmid. Restriction endonuclease digestion and ligation for gene fragment assembling and sub-cloning full length gene to expression vectors.

- Site-directed mutagenesis to make point mutation, deletion and insertion DNA constructs as well as green fluorescent protein (GFP) or Flag fusion PrP constructs for prion studies, and to repair

missing, extra insert, or miss-matched nucleotides in amplified gene fragments for ion channel

cloning projects.

• Cell biology and protein biochemistry:

- Ion channel gene expression by using traditional lipid-mediated or electroporation

transfection methods to deliver ion gene to CHO or HEK cells.

- In Vitro cell free system for RNA translation and protein synthesis analysis.

- Protein electrophoresis, Western blotting, Immunoprecipitation, immunochemistry, immunolocalization, antigen and antibody purification, Enzyme linked immunosorbent assay (ELISA). In vivo and in vitro labeling of proteins with radioactive isotopes and measuring protein degradation by pulse-chase analysis

- Enzyme kinetics, Inhibition studies, Spectroscopy (UV, visible, fluorescence)

• Cell death and apoptosis detection:

- Electron spin resonance (ESR) spectrum and CM-H2DCFDA dye for the detection of

reactive oxygen species.

- Measurement of products of lipid peroxidtion, TUNEL, DNA ladder gel electrophoresis,

Annexin-V binding for identification of apoptosis.

- Examination of Tyrosine kinase activities including ERK and p38MAPKase for signal

transduction studies.

• Cell and tissue culture:

Hands on culturing cardiac myocytes from neonatal rat, fibroblast cells from human skin, epithelial

cells from intestine (Caco-2 cells), neuron from human fetal brain, human or mouse neuroblastoma

cells, CHO, HEK, Hep 3B cells, hybridoma cells, and blood lymphocytes, Jurkat cells.

• Cell based ion channel assays by operating FLIPR and FLEXstation:

- Membrane potential assay

- Calcium Fluo-8 assay

- Calcium-4 assay

- Thallium FluxORTM potassium ion channel assay.

• Animal model development and study:

Experience in developing canine hemorrhagic shock and cardiac ischemic injury model, mouse

phenylketonuria (PKU) model, and mouse petroleum inhalation toxicity model. Carried out

tissue morphologic, biochemical metabolic, and physical functional studies with these animal models.

SELECTED PUBLICATIONS:

Das D, Luo X, Singh A, Gu Y, Ghosh S, Mukhopadhyay CK, Chen SG, Sy MS, Kong Q, Singh N (2010) Paradoxical role of prion protein aggregates in redox-iron induced toxicity. PLoS One 5(7):e11420.

Gu Y, Singh A, Bose S, Singh N. (2008) Pathogenic mutations in the glycosylphosphatidylinositol signal peptide of PrP modulate its topology in neuroblastoma cells. Mol Cell Neurosci. 37(4):647-56.

Gu Y, Verghese S, Bose S, Mohan M, Singh N. (2007) Mutant prion protein D202N associated with familial prion disease is retained in the endoplasmic reticulum and forms 'curly' intracellular aggregates. J Mol Neurosci.

32(1):90-6.

Gu Y, Luo X, Basu S, Fujioka H, Singh N. (2006) Cell-specific metabolism and pathogenesis of transmembrane prion protein. Mol Cell Biol. 26(7):2697-715.

Mishra RS, Basu S, Gu Y, Luo X, Zou WQ, Mishra R, Li R, Chen SG, Gambetti P, Fujioka H, Singh N.(2004) Protease-resistant human prion protein and ferritin are cotransported across Caco-2 epithelial cells: implications for species barrier in prion uptake from the intestine. J Neurosci. 24(50):11280-90.

Gu Y, Singh N. (2004) Doxycycline and protein folding agents rescue the abnormal phenotype of familial CJD H187R in a cell model. Brain Res Mol Brain Res. 123(1-2):37-44.

Gu Y, Hinnerwisch J, Fredricks R, Kalepu S, Mishra RS, Singh N. (2003) Identification of cryptic nuclear localization signals in the prion protein. Neurobiol Dis. 12(2):133-49.

Mishra RS, Bose S, Gu Y, Li R, Singh N. (2003) Aggresome formation by mutant prion proteins: the unfolding role of proteasomes in familial prion disorders. J Alzheimers Dis. 5(1):15-23.

Gu Y, Verghese S, Mishra RS, Xu X, Shi Y, Singh N. (2003) Mutant prion protein-mediated aggregation of normal prion protein in the endoplasmic reticulum: implications for prion propagation and neurotoxicity. J Neurochem. 84(1):10-22.

Mishra RS, Gu Y, Bose S, Verghese S, Kalepu S, Singh N. (2002) Cell surface accumulation of a truncated transmembrane prion protein in Gerstmann-Straussler-Scheinker disease P102L. J Biol Chem. 277(27):24554-61.

Singh N, Gu Y, Bose S, Kalepu S, Mishra RS, Verghese S. (2002) Prion peptide 106-126 as a model for prion replication and neurotoxicity. Front Biosci. 7:a60-71.

Gu Y, Fujioka H, Mishra RS, Li R, Singh N. (2002) Prion peptide 106-126 modulates the aggregation of cellular prion protein and induces the synthesis of potentially neurotoxic transmembrane PrP. J Biol Chem. 277(3):2275-86.

Gu Y, Jing Y, Kumar A, Sharma Y, Fujioka H, Singh N. (2001) Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126. J Alzheimers Dis. 3(2):169-180.

Gu Y, Zou Y, Aikawa R, Hayashic D, Kudoh S, Yamauchi T, Uozumi H, Zhu W, Kadowaki T, Yazaki Y and Komuro I. (2001) Growth hormone signalling and apoptosis in neonatal rat cardiomyocytes. Mol. Cell. Biochem. 223: 35-46.

Jin T, Gu Y, Zanusso G, Sy M, Kumar A, Cohen M, Gambetti P, Singh N. (2000) The chaperone protein BiP binds to a mutant prion protein and mediates its degradation by the proteasome. J Biol Chem. 275(49):38699-704.

Aikawa R, Nawano M, Gu Y, Katagiri H, Asano T, Zhu W, Nagai R, Komuro I. (2000) Insulin prevents cardiomyocytes from oxidative stress-induced apoptosis through activation of PI3 Kinase/Akt. Circulation 102(23); 2873-2879.

Yuan G and Gu Y. (1999) Epidermal growth factor receptor participates in growth hormone signaling pathway in cardiac myocytes of neonatal rat. Chin. Med. J. 112(6): 546-549.

Gu Y, Yuan G, and Lan Z. (1995) Study on pulmonary lipid peroxidation injury in rats induced by crude petroleum vapor inhalation. Chin. Public Health J. 14(4): 225-227.

Yuan G, Gu Y, Lan Z. (1995) Effect of crude petroleum vapor inhalation on anti-lipid- peroxidation

activities of blood and lung in rats. Chin. J. Naut. Med. 2(2): 18-21.

Gu Y, Yuan G, Ma J, Lan Z. (1993) A study on genetic toxicity of petroleum benzin with the method

of chromotest. Hygiene Toxicol. J. 7(4): 246-247.

Yuan G, Gu Y, Wang Y, Lan Z. (1993) A study on the lipid peroxidation injury of the rats by acute

petroleum vapor inhalation. Chin. labour hygiene and occupational Dis. J. 11(4): 203-205.

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